Plasticized bone grafts, and methods of making and using same

ABSTRACT

The present invention provides a plasticized dehydrated or freeze-dried bone and/or soft tissue product that does not require special conditions of storage, for example refrigeration or freezing, exhibits materials properties that approximate those properties present in normal hydrated tissue, is not brittle, does not necessitate rehydration prior to clinical implantation and is not a potential source for disease transmission. The invention replaces water in the molecular structure of the bone or soft tissue matrix with one or more plasticizers allowing for dehydration of the tissue, yet not resulting in an increase in brittleness of the plasticized product, and resulting in compressive and/or tensile properties similar to those of normal hydrated bone. Replacement of the chemical plasticizers by water prior to implantation is not required and thus, the dehydrated bone or soft tissue plasticized product can be placed directly into an implant site without significant preparation in the operating room.

FIELD OF THE INVENTION

[0001] The present invention provides a plasticized dehydrated boneand/or soft tissue product that does not require special conditions ofstorage, for example refrigeration or freezing, exhibits materialsproperties that approximate those properties present in normal hydratedtissue, is not brittle and does not necessitate rehydration prior toclinical implantation. The invention replaces water in the molecularstructure of the bone or soft tissue matrix with one or moreplasticizers allowing for dehydration of the tissue, yet not resultingin an increase in brittleness of the plasticized product, and resultingin compressive and/or tensile properties similar to those of normalhydrated bone. Replacement of the chemical plasticizers by water priorto implantation is not required and thus, the dehydrated bone or softtissue plasticized product can be placed directly into an implant sitewithout significant preparation in the operating room. The presentplasticized graft does not need rehydration, possesses adequatematerials properties, and is not a potential source for diseasetransmission.

BACKGROUND OF THE INVENTION

[0002] Bone tissue is a homogeneous material comprised of osteoid andminerals. The osteoid is a viscous gel-like material comprised primarilyof type I collagen (approximately 90%), proteoglycans, and varioussulfated and non-sulfated mucopolysaccharides. The mineral componentconsists primarily of a crystalline form of calcium phosphate, hydroxyapatite, with amounts of calcium carbonate, tricalcium phosphate, andsmaller amounts of other forms of mineral salts. This bone tissue islaid down around cells called osteocytes and these cells are found insmall interconnected channels (lacunnae) which are interconnectedthrough a series of channels comprising the Haversian canal system. Atthe level of the microscope, it is possible to observe that bone tissueis organized into osteons of compact bone made of concentric,perivascular layers of highly coaligned mineralized collagen fiberbundles. The predominant orientation within a single layer varies withrespect to the vascular axis and various combinations of orientation insuccessive lamellae result in variable overall collagen orientationwithin each osteon. Differences in overall collagen orientation aredirectly reflected in differing mechanical behavior of single osteons.Transversely oriented collagen results in better resistance tocompressive loading along the axis, whereas predominant longitudinalorientation results in better resistance to tensile stress. Thepredominant orientation of collagen within a cross-section of long boneis not random, but matches the expected distribution of mechanicalstress across the section, and its rotational shift along the wholeshaft. More transverse collagen is deposited at sites of compressiveloading, and more longitudinal collagen is deposited at sites of tensilestress. These structural oriented bone tissues in a load bearing boneare presumed to be laid down by the osteocytes present in the bone andbone remodeling mediates mechanical adaptation in compact bone.

[0003] A bone is typically comprised of bone tissue in the form ofcortical and trabecular bone. Cortical bone is frequently referred to ascompact bone and is the major load-bearing part of a bone. Trabecularbone is present in what is typically referred to as cancellous bonewhere it appears as a densely interconnected structure of “spongy” bone.Spongy bone in a typical bone contains the hemotopoietic cellularelements which is called bone marrow. Trabecular bone can be describedas forming a cross-bracing lattice between cortical bone in a bone. Itis important to emphasize a need to differentiate between “a bone” and“bone” (as a tissue). A bone is comprised of bone tissue present ascortical and cancellous (spongy) bone.

[0004] The mineralized osteoid typical of bone tissue is hydrated alongthe organic molecular structure and is an essential element of themineral structure. Hydrating molecules of water form complex molecularassociations with these organic and non-organic elements of bone tissueand can be described as being tightly bound, loosely bound, and free.Free water and loosely bound water can frequently be removed from bonetissue with only minor changes in the overall mechanical characteristicsof the bone tissue. Tightly bound water can be removed only underextreme conditions and results in significant changes in the physicaland mechanical properties of bone tissue. In fresh bone, water serves asolvating function in bone tissue allowing proper orientation andmolecular spacing of the collagen fibrils which maintain structuralalignment of the mineral phase in association with the organic phase.

[0005] Bone tissue in the form of bone grafts for implantation into apatient, is typically preserved and provided in a dehydrated state.Dehydration of bone tissue through drying, whether by air drying orsublimation as in freeze-drying, results in alteration of the molecularstructure of the bone tissue and as a result of the reorientation of thecollagen fibrils and the crystalline mineral phase, stress accumulatesin the bone tissue. This stress can be relieved by rehydration or by theoccurrence of small or large dislocations of structure. Smalldislocations are designated micro fractures and are not usually visibleto the naked eye. Large dislocations are designated fractures and areusually visible to the naked eye.

[0006] In a long bone, for example a femur, tibia, fibula, or humerus,the shaft separates the proximal and distal ends of the long bone. Theshaft serves to focus loads applied to the whole bone into a smallerdiameter than found at the proximal and distal ends of the long bone andthe shaft of a long bone is typically of a cylindrical shape and iscomprised of compact (cortical) bone. Loads applied along the axis ofthe shaft require that the cortical bone maintain a constantcircumference, i.e. the tendency to failure would distort the bonetissue perpendicular to the axis of load application. Thus, theorientation of the collagen fibers should be such that tensile stress isresisted along the axis of loading and compressive stress is resistedperpendicular to loading. Drying of shaft portions of long bones resultsin reorientation of collagen fibers and the mineral phase such thatchanges in the circumferential orientation create stress within the bonematrix which can be relieved only by rehydration or occurrence of afracture which allows a reorientation approximating the originalorientation. In dehydrated cortical ring grafts cut from shafts of longbones, this stress release can present as a fracture along the long axisof the bone shaft leaving a circumference which approximates thecircumference of the cortical ring graft prior to drying. By rehydratingbone grafts prior to implantation, the potential for fracture formationwhich can compromise the function of the bone product can be reduced,but not eliminated. Fractures as discussed above can occur in dehydratedbone prior to rehydration and result in a graft having compromisedbiomechanical properties, which in turn can result in graft failure whenimplanted in a patient.

[0007] Load-bearing soft tissue grafts such as ligaments, tendons, andfascia lata are frequently provided in a freeze-dried state. Such graftsmust be rehydrated prior to clinical implantation. Such soft tissuegrafts typically contain collagen, elastin, and assorted proteoglycansand mucopolysaccharides. The collagens and elastins are the load-bearingcomponent(s) of these soft tissue grafts and the assorted proteoglycansand polysaccharides serve to bind the fibrillar collagens into amatrix-like structure. The structural organization of fascia lata issimilar to dura mater in being somewhat isotropic in load-bearingproperties (Wolfinbarger, L, Zhang, Y, Adam, BLT, Homsi, D, Gates, K,and Sutherland, V, 1994, “Biomechanical aspects on rehydratedfreeze-dried human allograft dura mater tissues, J. AppliedBiomaterials, 5:265-270) whereas tendons (for example the Achillestendon) or ligaments (for example the Anterior cruciate ligament) aretypically anisotropic in load-bearing properties. In these types ofload-bearing soft tissue grafts, the tensile properties of the tissuesdepend on the flexibility of the collagenous structures to stretch underload and return to their original dimensions upon removal of the load.

[0008] A wide variety of bone and soft tissue products are used inveterinary, medical, orthopaedic, dental, and cosmetic surgeryapplications. These bone and soft tissue products can be used inload-bearing and non-load bearing applications and the bone and softtissue products can be supplied under a variety of forms. Bone productsare provided as fresh-frozen, freeze-dried, rehydrated freeze-dried,air-dried, organic solvent preserved, or provided preserved by othersimilar types of preservation methods. Each method of preservation ofbone products possesses selected advantages and disadvantages and thusthe method of preservation is generally modified to select for specificneeds of a given bone graft. Soft tissue products are typically providedas fresh-frozen or freeze-dried and each method of preservation of softtissue products possess selected advantages and disadvantages and thusthe method of preservation is generally modified to select for specificneeds of a given soft tissue product.

[0009] Bone and soft tissue products preserved and stored by methodsinvolving freeze-drying (removal of water by sublimation) yield a boneor soft tissue product which is significantly more brittle than normalbone and has a tendency to fracture into numerous small pieces, whichultimately can result in graft failure. Specifically, freeze-dryingcauses grafts to be brittle and typically causes shrinkage where theshrinkage is often not uniform, thereby causing graft failure; solventpreservation using for example, acetone or alcohol, can causeirreversible denaturation of proteins, and solubilization of solventsoluble components, including for example, lipids. These alterations inmaterials properties of the bone and soft tissue products necessitates arehydration step in preparation of the bone and soft tissue product forimplantation. However, rehydration does not solve the problem, graftscan fracture prior to rehydration, thereby making rehydration futile,and if there are micro fractures prior to rehydration they remain afterrehydration. These grafts are more likely to fail regardless of whetherthey are rehydrated. Even after rehydration the materials properties donot approximate the materials properties of normal bone.

[0010] Bone and soft tissue products are generally separated into loadbearing and non-load bearing products. Examples of non-load bearing boneproducts are ground demineralized bone which are used for inducing newbone formation in a particular implant site. Load-bearing bone productsare rarely demineralized and are used at implant sites where the bonegraft will be expected to withstand some level of physical load(s). Itis therefore important that load bearing bone products not fail duringnormal movement(s) of the implant recipient and that the bone productnot stimulate a pronounced physiological response. The majority of boneproducts are provided in either the fresh-frozen or freeze-dried format.The fresh-frozen format is undesirable because it includes donor derivedbone marrow and is thus immunogenic and a source of diseasetransmission. The freeze-dried format is less of a problem thanfresh-frozen grafts in the potential for disease transmission, however afreeze-dried bone graft is significantly more brittle than normal bone,more brittle than fresh frozen bone, and must be rehydrated prior toclinical usage. In that clinicians typically do not have time toadequately rehydrate bone graft products in the operating room, it isadvantageous to provide a dehydrated or freeze-dried bone product whichdoes not need rehydration, possesses adequate materials properties, andis not a potential source for disease transmission.

SUMMARY OF THE INVENTION

[0011] It is an objective of the present invention to provideimplantable, non-demineralized, load-bearing bone products which aremechanically stabilized in a dehydrated state by use of biocompatibleplasticizers.

[0012] It is a further objective of the present invention to provideimplantable, load-bearing, soft tissue products which are mechanicallystabilized in a dehydrated state by use of biocompatible plasticizers.

[0013] It is also an objective of the present invention to provideimplantable, load-bearing, bone products which do not requirerehydration.

[0014] It is yet a further objective of the present invention to provideimplantable, load-bearing, soft tissue products which do not requirerehydration.

[0015] It is an objective of the present invention to provide methods ofplasticizing load-bearing bone and soft tissue products.

[0016] It is a further objective of the present invention to provideplasticized bone and soft tissue products which are resistant toproliferation of microorganisms.

[0017] It is yet a further objective of the present invention to providebone and soft tissue products which can be stored at room temperatureusing conventional packaging.

[0018] It is a further objective of the present invention to provideplasticized bone and soft tissue products where the plasticizer can bereadily removed prior to implantation.

[0019] It is a further objective of the present invention to useplasticizers to plasticize bone and soft tissue products which are nottoxic to a recipient of the plasticized bone or soft tissue graft.

[0020] It is yet a further objective of the present invention to provideimplantable load-bearing bone and soft tissue products which are similarin physical, chemical, and biological properties as compared to normaltissue (fresh bone or fresh soft tissues) yet lack the inherentdisadvantages (including for example, potential disease transmission,increased immunogenicity, and a tissue (e.g. bone marrow) which canyield toxic degradation products and/or retard graft incorporation) offresh-frozen, dehydrated, and freeze-dried bone and/or soft tissueproducts.

[0021] It is a further objective of the present invention to provide aplasticized bone graft suitable for transplantation into a human,including a non-demineralized bone graft having an internal matrixessentially free from bone marrow elements; and one or more plasticizerscontained in the internal matrix.

[0022] It is an object of the present invention to provide a plasticizedbone graft, including a cleaned, non-demineralized, bone graft; and oneor more plasticizers, where the cleaned non-demineralized bone graft isimpregnated with the one or more plasticizers.

[0023] It is yet a further objective of the present invention to providea plasticized bone graft, including a cleaned, non-demineralized, bonegraft including one or more plasticizers.

[0024] It is a further objective of the present invention to provide amethod for producing a plasticized bone graft suitable fortransplantation into a human, by impregnating a cleaned,non-demineralized, bone graft with one or more plasticizers to produce aplasticized bone graft.

[0025] Plasticity of soft tissues depends primarily on the waters ofhydration present in the matrix structure, where water movement under aload is restricted by the viscous nature of theproteoglycan/polysaccharide component, and bound waters of hydration inthe collagen component affect the flexibility of the tensile componentof the tissue(s). The present invention deals with the plasticization ofthese load bearing tissue constructs where the water removed is replacedwith one or more plasticizers including for example, glycerol (glycerinUSP) (liquid substitution) such that the graft does not need to berehydrated or washed to remove the plasticizer prior to clinicalimplantation.

[0026] The present invention provides a dehydrated or freeze-driedplasticized bone or soft tissue product, preferably containing less than5% residual moisture, which product requires no or minimal processingjust prior to clinical implantation. The present invention solves priorart problems of grafts having insufficient materials properties, graftbrittleness, and the necessity for rehydration prior to clinicalimplantation, by providing a plasticized dehydrated bone and/or softtissue product that exhibits materials properties that approximate thoseproperties present in normal hydrated tissue, is not brittle and doesnot necessitate rehydration prior to implantation.

DETAILED DESCRIPTION

[0027] I. Definitions:

[0028] The below definitions serve to provide a clear and consistentunderstanding of the specification and claims, including the scope to begiven such terms.

[0029] Alcohol. By the term “alcohol” is intended for the purposes ofthe present invention, one of a series of organic chemical compounds inwhich a hydrogen attached to carbon is replaced by a hydroxyl. Suitablealcohols useful in the plasticizer composition of the present inventionpreferably include C₁-C₁₀ alcohols, and more preferably ethanol andisopropyl alcohol.

[0030] Allowash™ Solution. By the term “Allowash™ Solution” is intendedthose detergent compositions disclosed in co-pending U.S. patentapplication Ser. No. 08/620,856 incorporated herein by reference.Examples of suitable Allowash compositions include: a cleaningcomposition containing essentially about 0.06 wt %polyoxyethylene-4-lauryl ether, about 0.02 wt % poly (ethyleneglycol)-p-nonyl-phenyl-ether; about 0.02 wt % octyphenol-ethyleneoxideand endotoxin free deionized/distilled water.

[0031] Biocompatible. By the term “biocompatible” is intended for thepurposes of the present invention, any material which does not provokean adverse response in a patient. For example, a suitable biocompatiblematerial when introduced into a patient does not itself provoke asignificant immune response, and is not toxic to the patient.

[0032] Biomechanical strength. By the term “biomechanical strength” isintended for the purposes of the present invention, those propertiesexhibited by a tissue graft, including loading strength, compressivestrength, and tensile strength.

[0033] Bone graft. By the term “bone graft” is intended for the purposesof the present invention, any bone or piece thereof obtained from adonor for example a human or animal and/or cadaver donor, including forexample any essentially intact bone graft including for example thefemur, tibia, ilia, humorous, radius, ulna, ribs, whole vertebrae,mandibula and/or any bone which can be retrieved from a donor withminimal cutting of that bone for example, one half of an ulna, a femurcut in half to yield a proximal half and a distal half, femoral head,acetabula, distal femur, femur shaft, hemi-pelvi, humerus shaft,proximal femur, proximal femur with head, proximal humeri, proximaltibia, proximal tibia/plateaus, talus, tibia shaft, humeral head, ribs,and/or at least a substantial portion of a whole bone, i.e. at leastone-quarter of a whole bone; and/or any cut bone grafts including forexample an iliac crest wedge, a Cloward dowel, a cancellous cube, afibular strut, cancellous block, a crock dowel, femoral condyles,femoral ring, femur segment, fibula segment, fibular wedge, tibia wafer,ilium strip, Midas Rex dowel, tibial segment, and radius/ulna wedge.

[0034] Bone marrow elements. By the term “bone marrow elements” isintended for the purposes of the present invention, the highly cellularhematopoietic connective tissue filling the medullary cavities andspongy epiphysis of bones which may harbor bacterial and/or viralparticles and/or fungal particles, and includes for example, blood andlipid.

[0035] Cleaned bone graft. By the term “cleaned bone graft” is intendedfor the purposes of the present invention, a bone graft that has beenprocessed using means know in the art, to remove bone marrow elements.

[0036] Dehydrated bone or soft tissue. By the term “dehydrated bone orsoft tissue” is intended bone tissue or soft tissue which is preservedby dehydration, such drying methods including for example,freeze-drying, and/or sublimation and/or air drying and/or liquidsubstitution.

[0037] Essentially free from. By the term “essentially free from” isintended for the purposes of the present invention, a bone graft wherethe material removed (i.e., bone marrow elements) from the bone graft isnot detectable using detection means known in the art at the time offiling of this application.

[0038] Incubating. By the term “incubating” is intended for the purposesof the present invention, processing a bone graft in for example aplasticizer composition by soaking the graft in the composition, shakingthe graft with the composition, subjecting the graft to flow of thecomposition where the flow is induced by negative or positive pressure,subjecting the graft and/or the composition to negative or positivepressure, or soaking the bone graft in a plasticizer composition in anegative pressure environment.

[0039] Impregnating. By the term “impregnating” is intended for thepurposes of the present invention, any processing conditions whichresult in filling the internal matrix of a bone graft with a plasticizercomposition.

[0040] Internal matrix. By the term “internal matrix” is intended forthe purposes of the present invention, the spongy epiphysis of bones,the intercellular substance of bone tissue including collagen fibers andinorganic bone salts; or in soft tissue, the intercellular substance ofsuch soft tissue including for example ligaments and tendons, includingcollagen and elastin fibers and base matrix substances.

[0041] Load-bearing. By the term “load-bearing” is intended for thepurposes of the present invention a non-demineralized bone product orsoft tissue product for implantation in a patient at a site where thebone graft or soft tissue graft will be expected to withstand some levelof physical load(s).

[0042] Materials properties. By the term “materials properties” isintended for the purposes of the present invention, those propertiespresent in normal fresh bone which include for example, loadingstrength, compressive strength, tensile strength, and deformability.

[0043] Negative pressure. By the term “negative pressure” is intendedfor the purposes of the present invention, a pressure below atmosphericpressure, i.e. below 1 atm.

[0044] Normal bone or soft tissue. By the term “normal bone or softtissue” is intended for the purposes of the present invention, freshhydrated autogenous and/or fresh-frozen hydrated allograft tissueincluding for example, bone, fascia, ligaments, and tendons.

[0045] Permeation enhancer. By the term “permeation enhancer” isintended for the purposes of the present invention, any agent includingfor example, isopropyl alcohol, that facilitates penetration of the oneor more plasticizers or plasticizer composition into the bone or softtissue. In the case of isopropyl alcohol, permeation is enhanced due tothe reduced surface tension of the alcoholic solution.

[0046] Plasticization. By the term “plasticization” is intended for thepurposes of the present invention, replacing free and loosely boundwaters of hydration in a tissue(s) with one or more plasticizers withoutaltering the orientation of the collagen fibers and associated mineralphase.

[0047] Plasticizer. By the term “plasticizer” is intended for thepurposes of the present invention, any biocompatible compounds which aresoluble in water and can easily displace/replace water at the molecularlevel and preferably have a low molecular weight such that theplasticizer fits into the spaces available to water within the hydratedmolecular structure of the bone or soft tissue. Such plasticizers arepreferably not toxic to the cellular elements of tissue into which thegraft is to be placed, or alternatively, the plasticizer is easilyremoved from the graft product prior to implantation. Suitableplasticizers are preferably compatible with and preferably readilyassociates with the molecular elements of the bone tissue and/or softtissue. Suitable plasticizers include for example: glycerol (glycerinUSP), adonitol, sorbitol, ribitol, galactitol, D-galactose,1,3-dihydroxypropanol, ethylene glycol, triethylene glycol, propyleneglycol, glucose, sucrose, mannitol, xylitol, meso-erythritol, adipicacid, proline, hydroxyproline or similar water-soluble small molecularweight solutes which can be expected to replace water in the base matrixstructure of bone tissue and/or soft tissue and provide the hydratingfunctions of water in that tissue. Suitable solvents include forexample: water, alcohols, including for example ethanol and isopropylalcohol.

[0048] Plasticizer composition. By the term “plasticizer composition” isintended for the purposes of the present invention, any compositionwhich includes one or more plasticizers and one or more biocompatiblesolvents. Suitable solvents include for example: water, and alcohols,including for example C₁-C₁₀ alcohols, and more preferably ethanol andisopropyl alcohol.

[0049] Positive pressure. By the term “positive pressure” is intendedfor the purposes of the present invention, a pressure above atmosphericpressure, i.e. above 1 atm.

[0050] Rehydration. By the term “rehydration” is intended for thepurposes of the present invention, hydrating a dehydrated plasticizedtissue graft or a dehydrated non-plasticized tissue graft, with water,for example, prior to implantation into a patient. In the case of aplasticized graft, the plasticizer may optionally be not replaced bywater or may optionally be partially or fully replaced by water.

[0051] Soft tissue grafts. By the term “soft tissue grafts” is intendedfor the purposes of the present invention, load-bearing andnon-load-bearing soft tissue products. Non load-bearing grafts includecadaveric skin. Load-bearing soft tissue grafts include for example:pericardium, dura mater, fascia lata, and a variety of ligaments andtendons. Soft tissue grafts are composed of an internal matrix whichincludes collagen, elastin and high molecular weight solutes whereduring cleaning cellular elements and small molecular weight solutes areremoved.

[0052] II. Plasticizers

[0053] Plasticization of load-bearing bone or soft tissue graftsrepresents a method of replacing free and loosely bound waters ofhydration in the tissue(s) with a plasticizer composition containing oneor more plasticizers, without altering the orientation of the collagenfibers and associated mineral phase. Suitable plasticizers includecompounds which are soluble in water and can easily displace/replacewater at the molecular level. Suitable plasticizers preferably have alow molecular weight such that the plasticizer fits into the spacesavailable to water within the hydrated molecular structure of the boneor soft tissue. Such plasticizers are not toxic to the cellular elementsof tissue into which the graft is to be placed, or alternatively, theplasticizer is easily removed from the graft product prior toimplantation. Finally, the plasticizer is preferably compatible with andpreferably readily associates with the molecular elements of the bone orsoft tissue.

[0054] Plasticizers suitable for use in the present invention includefor example, a variety of biocompatible aqueous solutions. Examples ofacceptable plasticizers include, but are not restricted to, members ofthe polyol family (sugar alcohols) of compounds including C₂ to C₇polyols, monoglycerides (such as monoolein and monolinolein), andvarious short- and medium-chain free fatty acids (such short-chain freefatty acids preferably having a carbon chain length of less than sixC₆), and such medium-chain free fatty acids preferably having a carbonchain length of from C₁₂ to C₁₄) and their correspondingmonoacylglycerol esters (MGs) such as the saturated MGs, ranging incarbon chain length from C₅ to C₆, and preferably C₅ to C₁₄ MGs.Specific plasticizers include, but are not limited to, glycerol(glycerin USP), adonitol, sorbitol, ribitol, galactitol, D-galactose,1,3-dihydroxypropanol, ethylene glycol, triethylene glycol, propyleneglycol, glucose, sucrose, mannitol, xylitol, meso-erythritol, adipicacid, proline, hydroxyproline or similar water-soluble small molecularweight solutes which can be expected to replace water in the base matrixstructure of bone or soft tissue, and provide the hydrating functions ofwater in that tissue. Other plasticizers suitable for use in the presentinvention can be readily selected and employed by one of ordinary skillin the art to which the present invention pertains without undueexperimentation depending on the desired clinical outcome, sensitivityof the implantation procedure, patient sensitivities, and physicianchoice.

[0055] The present plasticizers are preferably employed at aconcentration in the range of from 0.1 to 2.0 M, 10% to 100% byweight/volume, or 3% to 30% by weight of bone or soft tissue. The use ofMolar concentrations and weight/volume percentages to express preferredconcentration ranges are intended to deal with the concentrations ofthese plasticizers in the solutions used to treat the tissues. The useof the weight percent of plasticizer in load-bearing bone or soft tissueis intended to deal with the effective quantity of a given plasticizerin the load-bearing tissue which is necessary to effectively replace thewaters of hydration present in the unprocessed tissues which aremaximally plasticized to a state approximating normal tissue. Theplasticizer can be introduced into the bone or soft tissue matrix at anynumber of steps in the processing procedures and at a variety ofconcentrations with and without the use of permeation enhancers.

[0056] The result(s) of plasticization of load-bearing bone and softtissue products are bone or soft tissue products which are similar totraditionally dehydrated bone and soft tissue products in residualmoisture but are not subject to fractures or micro fractures like suchdehydrated products, yet do not need to be rehydrated prior to use. Themechanical and use properties of a plasticized bone or soft tissueproduct are similar to those of natural (fresh autogenous and/orfresh-frozen allograft) bone, dura, pericardium, fascia, ligaments, andtendons.

[0057] III. Graft Cleaning and Processing

[0058] The present plasticizers may be introduced to the bone or softtissue products at several points in the processing procedure(s). Boneprocessing and cleaning procedures suitable for use with the presentinvention include known processes, as well as the processes described inU.S. Pat. No. 5,556,379 and co-pending U.S. patent application Ser. Nos.08/871,601 “Process for Cleaning Grafts Using Centrifugal Force and BoneGrafts Produced Thereby”; Ser. No. 08/620,858 “Composition for CleaningBones”; Ser. No. 08/646,520 “Recirculation Method for CleaningEssentially Intact Bone Grafts Using Pressure Mediated Flow of Solutionsand Bone Grafts Produced Thereby”; and Ser. No. 08/646,519 “UltrasonicCleaning of Allograft Bone” which are hereby incorporated herein intheir entirety. The plasticizers may be incorporated into the processingprocedure(s) using steps where the plasticizer(s) is/are present atessentially full strength, i.e. 100% concentration, in the presenceand/or absence of permeation enhancers, and at concentrations less thanfull strength.

[0059] Bone tissue is cleaned and processed as described in U.S. Pat.No. 5,556,379, and copending U.S. patent application Ser. Nos.08/871,601; 08/620,858; 08/646,520; and 08/646,519 by for example,transection of an essentially intact bone or perforation of anessentially intact bone with attachment of sterile plastic tubing to thecut end of a transected bone or to an attachment port inserted into theperforation of the perforated bone. The bone is immersed in a cleaningsolution, such solutions including known cleaning agents as well asthose described in the above-identified patent and co-pending patentapplications, with or without use of sonication. The cleaning solutionis induced to flow into, through, and out of the bone through use of aperistaltic pump or negative pressure applied to the cleaning solution,The induced flow of cleaning solution draws the bone marrow from theinterior of the bone, and particularly from the cancellous bone marrowspace, where it can be safely deposited in a receiving containercontaining a strong virucidal agent such as sodium hypochlorite (commonbleach). The cleaned bone can then be further cleaned by causing thecleaning solution to be replaced with a solution of one or moredecontaminating agents, including for example 3% hydrogen peroxide, withor without plasticizer. Hydrogen peroxide which in addition to its milddisinfection activity generates oxygen bubbles that can further assistin dislodging residual bone marrow materials causing the residual bonemarrow materials to flow from the bone and into the receiving container.

[0060] In the above-described process, after processing with thecleaning solution, after processing with a decontaminating agent, inplace of processing with a decontaminating agent, or after dehydration,the cleaned graft is plasticized for example, by processing the cleanedgraft with a plasticizer composition containing one or more plasticizersincluding for example glycerin USP in a solvent.

[0061] IV. Plasticization

[0062] Bone and soft tissue grafts can be cleaned and processed usingconventional methods. including those described in. When processingusing these methods the graft is plasticized by adding one or moreplasticizers or a plasticizer composition to processing steps after bonecleaning is essentially completed, and prior to freeze-drying. Underfreeze-drying, the water present in the bone (or smaller cut bone graftsproduced form the essentially intact bone) is removed by sublimation,however, the glycerol will remain and replace the free and bound wateras the water is removed from the bone tissue. The one or moreplasticizer(s) is added to fully hydrated bone tissue and theplasticizer(s) are induced to penetrate into the bone tissue optionallyusing a permeation enhancer. Thus, the bone or soft tissue is dehydratedyet the materials properties of the bone tissue will be similar to thematerials properties of normal bone or soft tissue, i.e. partially orfully hydrated bone or soft tissue. The produced plasticized bone orsoft graft contains minimal quantities of the plasticizer(s) and can beremoved from the package and directly implanted into a patient withoutrehydration. If the presence of these small quantities of glycerol is ofconcern, the bone or soft tissue grafts may be quickly rinsed and/orwashed in sterile saline just prior to implantation.

[0063] Bone or soft tissue cleaned and processed by the methods asdescribed for bone cleaning and processing in U.S. Pat. No. 5,556,379,and/or co-pending U.S. patent application Ser. Nos. 08/871,601;08/620,858; 08/646,520; and/or 08/646,519 and/or bone or soft tissuecleaned and processed by conventional methods, may be plasticized byprocessing with the plasticizer composition containing one or moreplasticizers, including for example glycerin USP, in a solvent by forexample drawing the plasticizer composition into the bone. Suitablesolvents include for example, 70% isopropyl alcohol. The 70% isopropylalcohol/plasticizer composition can be prepared by diluting absolute(100%) isopropyl alcohol with the one or more plasticizers, includingfor example glycerin USP such that the plasticizer accounts for 30% ofthe total volume and isopropyl alcohol accounts for 70% of the totalvolume. Under this method, the original processing procedures asdescribed in U.S. Pat. No. 5,556,379 regarding the use of 70% isopropylalcohol, is retained essentially unchanged. The isopropyl alcoholfacilitates penetration of the glycerol into the tissue by acting as apermeation enhancer and the glycerol more readily penetrates the tissuedue to the reduced surface tension of the alcoholic solution. Theinduced flow of glycerol/isopropyl alcohol into, through, and out of forexample, the essentially intact bone, further serves to remove residualcellular elements, for example bone marrow materials, if any. It alsoallows penetration of the glycerol/isopropyl alcohol solution into themost remote areas of the tissue, and facilitates a uniform distributionof the glycerol into the tissue. The isopropyl alcohol can be removedfrom the tissue by washing with a washing solution including sterilewater, for example as described in U.S. Pat. No. 5,556,379 following thealcohol processing step. Preferably, the washing solution includesglycerin USP (30% volume:volume). The washing solution facilitatesremoval of the isopropyl alcohol without removal of the glycerin USP.The cleaned and plasticized tissue can then be frozen and freeze-driedor dehydrated according to standard protocols.

[0064] Alternatively, bone or soft tissue grafts may be plasticizedafter cleaning and freeze-drying. For example, tissue can be processedand cleaned according to any method including known methods, or asdescribed in U.S. Pat. No. 5,556,379 described above. After the sterilewater wash the tissue (for example bone tissue) is cleaned of virtuallyall cellular elements (for example, bone marrow) present in the tissueand the cleaned tissue can be further processed into for example, smallcut bone grafts, and dehydrated or freeze-dried (also calledlyophilized) using standard methods well known to those skilled in theart. Freeze-dried or dehydrated tissue grafts preferably contain lessthan about 5% residual moisture, satisfying the definition offreeze-dried bone allografts as prescribed under Standards of theAmerican Association of Tissue Banks.

[0065] Clean freeze-dried or dehydrated bone or soft tissue grafts areplasticized by processing the tissue graft with a plasticizercomposition, suitable compositions including for example 70% isopropylalcohol/30% glycerin USP or 100% glycerin USP. Due to the presence ofair in the cancellous and cortical bone spaces, the plasticizer(s) mayonly penetrate into the bone tissue with which it is in physicalcontact. Suitable methods for achieving physical contact between theplasticizer and bone or soft tissue include those methods known to oneof ordinary skill in the art to which the present invention pertains.The plasticizer composition can be induced to flow into the cancellousand cortical bone spaces of bone tissue, or soft tissue, thus achievingphysical contact, by various known methods that can be readily selectedand employed by one of ordinary skill in the art to which the presentinvention pertains without undue experimentation, and include forexample, agitation of the a tissue with the plasticizer composition,application of a vacuum (5 to 500 mTorr) above the plasticizer. Thevacuum induces the air trapped in the, for example cancellous andcortical bone spaces/tissue to exit and be carried off. As the trappedair is removed from the cancellous and cortical bone spaces/tissue, theplasticizer quickly moves into the spaces previously occupied by airgreatly enhancing penetration of the plasticizer into the bone or softtissue. The plasticizer fills the spaces previously occupied by the freeand bound water restoring the tissue to a materials property similar tothat materials property of the original fully or partially hydratedtissue (e.g. normal bone).

[0066] The present one or more plasticizers may be introduced to softtissue products at several points in the processing procedures, but arepreferably introduced prior to the freeze-drying or dehydrating step. Byintroducing plasticizers prior to freeze-drying or dehydrating, thederived soft tissue graft is in a freeze-dried/dehydrated state wherethe plasticizer is used to stabilize the matrix and load bearingcomponents of the soft tissue graft such that the graft can be usedwithout rehydration/reconstitution.

[0067] V. Transplantation into a Patient

[0068] Prior to transplantation into a patient, excess glycerol mayoptionally be removed from the plasticized bone or soft tissue graftusing for example, the method described in co-pending U.S. patentapplication Ser. No. 08/871,601. Specifically, the plasticized graftsare placed into centrifuge vessels/containers and on top of insertsdesigned to keep the bone grafts off of the bottom of the containers.The grafts are then centrifuged at 1,000 to 2,000 revolutions per minute(rpm) for 10-20 minutes. The excess glycerol or similar plasticizerexits the grafts and collects in the bottom of the centrifuge containersaway from the grafts. The plasticizer tightly associated with themolecular and chemical structure of the tissue will not exit the graftand the tissue will remain plasticized without retaining physicallydiscemable quantities of plasticizer. The plasticized graft(s) may thenbe packaged directly or packaged in a packaging format which permitsapplication of a vacuum to the container. The current value of using apackaging format which permits storage of grafts under vacuum lies inthe ability to predict possible loss of sterility with loss of vacuum tothe packaging.

[0069] Clinical usage of plasticized bone or soft tissue grafts includesdirect implantation of the grafts without further processing followingremoval from the packaging, implantation following a brief washing insterile isotonic saline to remove any remaining traces of plasticizerassociated with the immediate surfaces of the grafts, or by implantationfollowing an extended (approximately 1 hour) washing with sterileisotonic saline to remove as much plasticizer as possible. Under any ofthe above described further processing of grafts, the materialsproperties of the plasticized grafts resemble those materials propertiesof fully or partially hydrated natural tissue (i.e. normal bone or softtissue). The produced plasticized graft does not need to be rehydratedprior to clinical implantation, yet retains the strength andcompressive/tensile properties of natural tissue. Plasticizedfreeze-dried soft tissue grafts where the plasticizer is used tostabilize the matrix and load bearing components of the soft tissuegraft, can also be directly implanted in a patient withoutrehydration/reconstitution.

[0070] Suitable surgical methods for implanting bone and soft tissuegrafts into a patient are well known to those of ordinary skill in theart to which the present invention pertains, and such methods areequally applicable to implantation of the present plasticized grafts.Those of ordinary skill in the art to which the present inventionpertains can readily determine, select and employ suitable surgicalmethods without undue experimentation.

[0071] Further details of the process of the invention are presented inthe examples that follow:

EXAMPLE 1 Processing of a Frozen Distal Femur

[0072] A. Cleaning and Processing:

[0073] A frozen distal femur is selected and all of the soft tissue andperiosteum is removed using sharp dissection techniques and periostealelevators. The graft is then transected to the desired length using aStryker® saw or band saw. Each bisected piece is not more than 30 cm inlength and is straight and contains no bone fragments. The surfacecartilage is then removed from the fem oral condyle with either ascalpel blade, periosteal elevator, or osteotome. The processinginstructions dictate leaving the cartilage “on” when appropriate. Usinga ⅜″ drill bit, the cut end of the shaft is drilled approximately 5 cm.The interior of the intramedullary canal is then throughly washed withthe lavage system.

[0074] An intercalary fitting is then inserted by screwing the threaded,tapered end into the cut end of the graft. The vacuum tubing isassembled by securing one end of the tubing to the nipple end of theintercalary fitting. The other end of the tubing is secured to thepiston driven pump. Finally, another section of vacuum tubing is securedto the other side of the piston pump. Approximately 4000 cc of a 1:100dilution of the “Allowash™ Solution” is poured into the sterile flushingvessel. The “Allowash™ Solution” is prepared by adding 4 cc of cleaningreagent to 3996 cc of sterile water. The flushing vessel is labeled as“Allowash™ Solution.” The open end of the second piece of vacuum tubingis placed into a graduated flask. The piston pump is set to “reverse”and the flow rate controller is set to 50%. The pump is turned on and atleast 500 cc of the first solvent (Allowash™ Solution) is drawn towaste. Thereafter, the open end of the second piece of vacuum tubing isremoved from the graduated flask and placed into the sterile flushingvessel. The drive is maintained in the “reverse” position at 50%. TheAllowash Solution recirculates for a minimum of 15 minutes.

[0075] The 1:100 dilution of the Allowash™ Solution is then decanted andapproximately 4 liters of 3% hydrogen peroxide is added to the flushingvessel. The piston pump is set to reverse and the flow rate controlleris set to 50%. The pump is then turned on and at least 500 cc of the 3%hydrogen peroxide solution is drawn to waste. Thereafter, the open endof the second piece of vacuum tubing is removed from the graduated flaskand placed it into the sterile flushing vessel. The drive is maintainedin the reverse position at 50%. The hydrogen peroxide is then allowed torecirculate for a minimum of 15 minutes.

[0076] The hydrogen peroxide is then decanted and approximately 3980 ccof sterile water is added along with the entire contents ofreconstituted vials of Bacitracin and Polymyxin B to the flushingvessel. The flushing vessel is clearly labeled “antibiotic.” The pistonpump is then set to reverse and the flow rate controller is set at 50%.The pump is turned on and at least 500 cc of antibiotic solution isdrawn to waste. The open end of the second piece of vacuum tubing isremoved from the graduated flask and placed into the sterile flushingvessel. The drive is maintained in the reverse position at 50%. Theantibiotic solution is allowed to recirculate for a minimum of 15minutes.

[0077] B. Plasticization:

[0078] The antibiotic solution is then decanted and approximately 4liters of 70% isopropyl alcohol/30% glycerin USP is added to theflushing vessel. The flushing vessel is clearly labeled as 70% IPA/30%glycerin USP. The piston pump is set to reverse and the flow ratecontroller is set to 50%. The pump is turned on and at least 500 cc ofIPA/glycerin USP solution is drawn to waste.

[0079] The open end of the second piece of vacuum tubing is removed fromthe graduated flask and placed into the sterile flushing vessel. Thedrive is maintained in the reverse position and the flow controller isset to 50%. The IPA/glycerin USP is allowed to recirculate for a minimumof 30 minutes. The IPA/glycerin USP solution is decanted and 4 liters of30% glycerin USP in sterile water is added to the flushing vessel. Theflushing vessel is labeled as glycerin USP washing solution. The pistonpump is set to reverse and the flow rate controller is set to 50%. Thepump is turned on and at least 500 cc of washing solution is drawn towaste.

[0080] The open end of the second piece of vacuum tubing is removed fromthe graduated flask and placed into the sterile flushing vessel. Thedrive is maintained in the reverse position and the flow rate controlleris set to 50%. The washing solution is allowed to recirculate for aminimum of 15 minutes. Thereafter, the bone graft is removed from theflushing vessel and processed for freeze-drying as per standardoperating procedure.

EXAMPLE 2 Processing of a Frozen Distal Femur

[0081] A. Cleaning and Processing:

[0082] A frozen distal femur is selected and all of the soft tissue andperiosteum is removed using sharp dissection techniques and periostealelevators. The graft is then transected to the desired length using aStryker® saw or band saw. Each bisected piece is not more than 30 cm inlength and is straight and contains no bone fragments. The surfacecartilage is then removed from the femoral condyle with either a scalpelblade, periosteal elevator, or osteotome. The processing instructionsdictate leaving the cartilage “on” when appropriate. Using a ⅜″ drillbit, the cut end of the shaft is drilled approximately 5 cm. Theinterior of the intramedullary canal is then throughly washed with thelavage system.

[0083] An intercalary fitting is then inserted by screwing the threaded,tapered end into the cut end of the graft. The vacuum tubing isassembled by securing one end of the tubing to the nipple end of theintercalary fitting. The other end of the tubing is secured to thepiston driven pump. Finally, another section of vacuum tubing is securedto the other side of the piston pump. Approximately 4000 cc of a 1:100dilution of the “Allowash™ Solution” is poured into the sterile flushingvessel. The “Allowash™ Solution” is prepared by adding 4 cc of cleaningreagent The flushing vessel is labeled as “Allowash™ Solution.” The openend of the second piece of vacuum tubing is placed into graduated flask.The piston pump is set to “reverse” and the flow rate controller is setto 50%. The pump is turned on and at least 500 cc of the first solvent(Allowash Solution) is drawn to waste. Thereafter, the open end of thesecond piece of vacuum tubing is removed from the graduated flask andplaced into the sterile flushing vessel. The drive is maintained in the“reverse” position at 50%. The Allowash Solution recirculates for aminimum of 15 minutes.

[0084] B. Plasticization:

[0085] The 1:100 dilution of the Allowash™ Solution is decanted andapproximately 4 liters of 3% hydrogen peroxide/30% glycerin USP is addedto the flushing vessel. The piston pump is set to reverse and the flowrate controller is set to 50%. The pump is turned on and at least 500 ccof the 3% hydrogen peroxide/glycerin USP solution is drawn to waste. Theopen end of the second piece of vacuum tubing is removed from thegraduated flask and placed into the sterile flushing vessel. The driveis maintained in the reverse position. The hydrogen peroxide/glycerinUSP is allowed to recirculate for a minimum of 15 minutes.

[0086] The hydrogen peroxide/glycerin USP is then decanted andapproximately 3980 cc of sterile water is added along with the entirecontents of reconstituted vials of Bacitracin and Polymyxin B preparedin a water solution of 30% glycerin USP, to the flushing vessel. Theflushing vessel is clearly labeled “antibiotic.” The piston pump is thenset to reverse and the flow rate controller is set at 50%. The pump isturned on and at least 500 cc of antibiotic solution is drawn to waste.The open end of the second piece of vacuum tubing is removed from thegraduated flask and placed into the sterile flushing vessel. The driveis maintained in the reverse position at 50%. The antibiotic solution isallowed to recirculate for a minimum of 15 minutes.

[0087] The antibiotic solution is then decanted and approximately 4liters of 70% isopropyl alcohol/30% glycerin USP is added to theflushing vessel. The flushing vessel is clearly labeled as 70% IPA/30%glycerin USP. The piston pump is set to reverse and the flow ratecontroller is set to 50%. The pump is turned on and at least 500 cc ofIPA/glycerin USP solution is drawn to waste.

[0088] The open end of the second piece of vacuum tubing is removed fromthe graduated flask and placed into the sterile flushing vessel. Thedrive is maintained in the reverse position and the flow controller isset to 50%. The IPA/glycerin USP is allowed to recirculate for a minimumof 30 minutes. The IPA/glycerin USP solution is decanted and 4 liters of30% glycerin USP in sterile water is added to the flushing vessel. Theflushing vessel is labeled as glycerin USP washing solution. The pistonpump is set to reverse and the flow rate controller is set to 50%. Thepump is turned on and at least 500 cc of washing solution is drawn towaste.

[0089] The open end of the second piece of vacuum tubing is removed fromthe graduated flask and placed into the sterile flushing vessel. Thedrive is maintained in the reverse position and the flow rate controlleris set to 50%. The washing solution is allowed to recirculate for aminimum of 15 minutes. Thereafter, the bone graft is removed from theflushing vessel and processed for freeze-drying as per standardoperating procedure.

EXAMPLE 3 Processing of a Frozen Distal Femur

[0090] A. Cleaning and Processing:

[0091] A frozen distal femur is selected and all of the soft tissue andperiosteum is removed using sharp dissection techniques and periostealelevators. The graft is then transected to the desired length using aStryker® saw or band saw. Each bisected piece is not more than 30 cm inlength and is straight and contains no bone fragments. The surfacecartilage is then removed from the femoral condyle with either a scalpelblade, periosteal elevator, or osteotome. The processing instructionsdictate leaving the cartilage “on” when appropriate. Using a ⅜″ drillbit, the cut end of the shaft is drilled approximately 5 cm. Theinterior of the intramedullary canal is then throughly washed with thelavage system.

[0092] An intercalary fitting is then inserted by screwing the threaded,tapered end into the cut end of the graft. The vacuum tubing isassembled by securing one end of the tubing to the nipple end of theintercalary fitting. The other end of the tubing is secured to thepiston driven pump. Finally, another section of vacuum tubing is securedto the other side of the piston pump. Approximately 4000 cc of a 1:100dilution of the “Allowash™ Solution” is poured into the sterile flushingvessel. The “Allowash™ Solution” is prepared by adding 4 cc of cleaningreagent. The flushing vessel is labeled as “Allowash™ Solution.” Theopen end of the second piece of vacuum tubing is placed into a graduatedflask. The piston pump is set to “reverse” and the flow rate controlleris set to 50%. The pump is turned on and at least 500 cc of the firstsolvent (Allowash Solution) is drawn to waste. Thereafter, the open endof the second piece of vacuum tubing is removed from the graduated flaskand placed into the sterile flushing vessel. The drive is maintained inthe “reverse” position at 50%. The Allowash Solution recirculates for aminimum of 15 minutes.

[0093] The 1:100 dilution of the Allowash™ Solution is then decanted andapproximately 4 liters of 3% hydrogen peroxide is added to the flushingvessel. The piston pump is set to reverse and the flow rate controlleris set to 50%. The pump is then turned on and at least 500 cc of the 3%hydrogen peroxide solution is drawn to waste. Thereafter, the open endof the second piece of vacuum tubing is removed from the graduated flaskand placed it into the sterile flushing vessel. The drive is maintainedin the reverse position at 50%. The hydrogen peroxide is then allowed torecirculate for a minimum of 15 minutes.

[0094] The hydrogen peroxide is then decanted and approximately 3980 ccof sterile water is added along with the entire contents ofreconstituted vials of Bacitracin and Polymyxin B to the flushingvessel. The flushing vessel is clearly labeled “antibiotic.” The pistonpump is then set to reverse and the flow rate controller is set at 50%.The pump is turned on and at least 500 cc of antibiotic solution isdrawn to waste. The open end of the second piece of vacuum tubing isremoved from the graduated flask and placed into the sterile flushingvessel. The drive is maintained in the reverse position at 50%. Theantibiotic solution is allowed to recirculate for a minimum of 15minutes.

[0095] The antibiotic solution is then decanted and approximately 4liters of 70% isopropyl alcohol (IPA) is added to the flushing vessel.The flushing vessel is labeled as 70% IPA. The piston pump is set toreverse and the flow rate controller is set to 50%. The pump is turnedon and at least 500 cc of IPA solution is drawn to waste. The open endof the second piece of vacuum tubing is removed from the graduated flaskand placed into the sterile flushing vessel. The drive is maintained inthe reverse position and the flow controller is set to 50%. The IPArecirculates for a minimum of 15 minutes. The IPA solution is thendecanted and 4 liters of sterile water is added to the flushing vessel.The flushing vessel is labeled as “washing solution.” The piston pump isset to reverse and the flow rate controller is set to 50%. The pump isturned on and at least 500 cc of washing solution is drawn to waste.

[0096] The open end of the second piece of vacuum tubing is removed fromthe graduated flask and placed into the sterile flushing vessel. Thedrive is maintained in the reverse position and the flow rate controlleris set to 50%. The washing solution recirculates for a minimum of 15minutes. The bone graft is removed from the flushing vessel andprocessed for freeze-drying as per standard operating procedure.

[0097] B. Plasticization:

[0098] The freeze-dried bone graft(s) are then placed into sterileglycerin USP such that they are totally immersed in the viscousglycerol. Vacuum (10 to 500 mTorr, preferably 100 to 200 mTorr) isapplied to the container until bubbles cease to exit the bone graft(about 5 to 60 minutes depending on the size and configuration of thebone graft, preferably about 20 to 30 minutes). The bone graft(s) arethen removed from the glycerin USP solution and placed into anappropriate centrifuge container on top of a graft support.

[0099] The bone graft(s) are centrifuged at about 1000 to 2000 rpm untilthe glycerol ceases to exit the bone graft and accumulate in the bottomof the centrifuge container (usually 5 to 60 minutes depending on thesize and configuration of the bone graft, preferably about 5 to 15minutes). The bone graft(s) are then removed from their respectivecentrifuge containers and packaged for distribution.

EXAMPLE 4 Processing Cloward Dowels

[0100] A. Cleaning and Processing:

[0101] Graft material is selected and all of the soft tissue andperiosteum is removed from the distal femur, proximal and distal tibia,and cartilage is removed from the site. The femur is transected 10-15 cmabove the femoral condyles and the distal femoral condyles are bisected.Transect the proximal tibia 10-15 cm below the tibial plateau. Thedistal femur or proximal tibia is placed in a Pan-A-Vise™. This isaccomplished by removing a section of the diaphysis, allowing the visejaws to grip the tissue securely. The Cloward set (12, 14,16, 18, or 20mm) is then assembled: 1. Place the extractor assembly within the cuttershaft, 2. Screw the cutter assembly onto the shaft with the aid of theCloward set wrench, 3. Screw the set-point onto the extractor assembly,4. Insert the shaft of the Cloward set into the ⅜″ variable speed drilland tighten the chuck with the key. The set-point is then placed andlocked at the forward aspect of the cutter.

[0102] The apparatus is then placed on the tissue to be fashioned.Drilling is commenced at a moderate speed. After the set-point has madea deep cut in the tissue, and the teeth have begun to cut into thetissue, drilling is stopped, and the set-point apparatus is unlocked.Drilling is continued using the marks created as a guide.

[0103] The Cloward(s) are then removed from the tissue block. A Stryker®saw or band saw is then used to remove the cut grafts after all havebeen cut. Any cartilage is then trimmed from the cortical face of theCloward(s) using a scalpel and a # 10 blade. The distal end of the graftis then trimmed perpendicular to the body of the graft with a band sawmaking sure the fashioned graft is at least 15 mm long. The Cloward(s)are cleansed using pulsatile water apparatus. If the surface marrow isnot easily removed, dry spin the graft(s) at 2600 rpm for 3 minutes.

[0104] The Cloward(s) are then placed in a sterile container withhydrogen peroxide (3%) at 37 to 44° C., The container is sealed and thecontainer is placed into the centrifuge. The centrifuge is thenbalanced. The grafts are then centrifuged at 2600 rpm for 15 minutes.The tissue is removed from the centrifuge and the grafts are placed intoan ultrasonic cleaner. Equal volumes of Allowash™ Solution, hydrogenperoxide (3%), and antibiotics are added to the ultrasonic cleaner andsonicate the tissue at 37-44° C. for a minimum of 1 hour. Thereafter,the tissue is removed from the ultrasonic cleaner.

[0105] The mixture is decanted and a sterile glass container is filledwith fresh 3% hydrogen peroxide. The grafts are then placed in thecontainer, the top is sealed and the container is taken to the largeultrasonic cleaner. The grafts are then sonicated for 90 minutes.Thereafter, the grafts are incubated overnight at 37-44° C. (minimum of6 hours, preferably 12 to 18 hours).

[0106] B. Plasticization:

[0107] After incubation, the hydrogen peroxide is decanted and the basinis filled with 70% isopropyl alcohol/30% glycerin USP and the grafts areincubated at room temperature for a minimum of 30 minutes. Thereafter,the isopropyl alcohouglycerin USP solution is decanted and the containeris filled with warm 30% glycerin USP in water. The grafts are incubatedfor a minimum of 30 minutes. Methods of incubation include for example:soaking.

[0108] The glycerin solution is then decanted and the Cloward dowels areremoved from the container. The Cloward dowels are then placed into asterile container. The container is sealed and placed into thecentrifuge. The centrifuge is balanced and the grafts are centrifugedfor 3-5 minutes to dry, and the remaining solution is removed.

[0109] The width and length of the Cloward(s) are measured, graftidentification numbers are assigned, and the information is recorded onthe “Tissue Processing Log Worksheet”. One graft is then placed into aglass, 120 cc bottle and the printed label is affixed with the uniquenumeric designator. This step is repeated until all deposits arebottled. The bottled grafts are either frozen and packaged, or frozenand freeze-dried and packaged.

EXAMPLE 5 Processing Cloward Dowels

[0110] A. Cleaning and Processing:

[0111] Graft material is selected and all of the soft tissue andperiosteum is removed from the distal femur, proximal and distal tibia,and cartilage is removed from the site. The femur is transected 10-15 cmabove the femoral condyles and the distal femoral condyles are bisected.Transect the proximal tibia 10-15 cm below the tibial plateau. Thedistal femur or proximal tibia is placed in a Pan-A-Vise™. This isaccomplished by removing a section of the diaphysis, allowing the visejaws to grip the tissue securely. The Cloward set (12, 14, 16, 18, or 20mm) is then assembled: 1. Place the extractor assembly within the cuttershaft, 2. Screw the cutter assembly onto the shaft with the aid of theCloward set wrench, 3. Screw the set-point onto the extractor assembly,4. Insert the shaft of the Cloward set into the ⅜″ variable speed drilland tighten the chuck with the key. The set-point is then placed andlocked at the forward aspect of the cutter.

[0112] The apparatus is then placed on the tissue to be fashioned.Drilling is commenced at a moderate speed. After the set-point has madea deep cut in the tissue, and the teeth have begun to cut into thetissue, drilling is stopped, and the set-point apparatus is unlocked.Drilling is continued using the marks created as a guide.

[0113] The Cloward(s) are then removed from the tissue block. A Stryker®saw or band saw is then used to remove the cut grafts after all havebeen cut. Any cartilage is then trimmed from the cortical face of theCloward(s) using a scalpel and a # 10 blade. The distal end of the graftis then trimmed perpendicular to the body of the graft with a band sawmaking sure the fashioned graft is at least 15 mm long. The Cloward(s)are cleansed using pulsatile water apparatus. If the surface marrow isnot easily removed, dry spin the graft(s) at 2600 rpm for 3 minutes.

[0114] B. Plasticization:

[0115] The Cloward(s) are then placed in a sterile container withhydrogen peroxide (3%) and glycerin USP (30%) at 37 to 44° C. Thecontainer is sealed and the container is placed into the Centrifuge. Thecentrifuge is then balanced. The grafts are then centrifuged at 2600 rpmfor 15 minutes. The tissue is removed from the centrifuge and the graftsare placed into an ultrasonic cleaner. Equal volumes of Allowash™Solution, hydrogen peroxide (3%), 30% glycerin USP, and antibiotics areadded to the ultrasonic cleaner and the tissue is sonicated at 37-44° C.for a minimum of 1 hour. Thereafter, the tissue is removed from theultrasonic cleaner.

[0116] The mixture is decanted and a sterile glass container is filledwith fresh 3% hydrogen peroxide/30% glycerin USP. The grafts are thenplaced in the container, the top is sealed and the container is taken tothe large ultrasonic cleaner. The grafts are then sonicated for 90minutes. Thereafter, the grafts are incubated overnight at 37-44° C.(minimum of 6 hours, preferably 12 to 18 hours).

[0117] After incubation, the hydrogen peroxide is decanted and the basinis filled with 70% isopropyl alcohol/30% glycerin USP and the grafts areincubated at room temperature for a minimum of 30 minutes. Thereafter,the isopropyl alcohol/glycerin USP solution is decanted and thecontainer is filled with warm 30% glycerin USP in water. The grafts areincubated for a minimum of 30 minutes. Methods of incubation include forexample: soaking and mild agitation.

[0118] The glycerin solution is then decanted and the Cloward dowels areremoved from the container. The Cloward dowels are then placed into asterile container. The container is sealed and placed into thecentrifuge. The centrifuge is balanced and the grafts are centrifugedfor 3-5 minutes to dry, and the remaining solution is removed.

[0119] The width and length of the Cloward(s) are measured, graftidentification numbers are assigned, and the information is recorded onthe “Tissue Processing Log Worksheet”. One graft is then placed into aglass, 120 cc bottle and the printed label is affixed with the uniquenumeric designator. This step is repeated until all deposits arebottled. The bottled grafts are either frozen and packaged, or frozenand freeze-dried and packaged.

EXAMPLE 6 Processing Cloward Dowels

[0120] A. Cleaning and Processing:

[0121] Graft material is selected and all of the soft tissue andperiosteum is removed from the distal femur, proximal and distal tibia,and cartilage is removed from the site. The femur is transected 10-15 cmabove the femoral condyles and the distal femoral condyles are bisected.Transect the proximal tibia 10-15 cm below the tibial plateau. Thedistal femur or proximal tibia is placed in a Pan-A-Vise™. This isaccomplished by removing a section of the diaphysis, allowing the visejaws to grip the tissue securely. The Cloward set (12, 14, 16, 18, or 20mm) is then assembled: 1. Place the extractor assembly within the cuttershaft, 2. Screw the cutter assembly onto the shaft with the aid of theCloward set wrench, 3. Screw the set-point onto the extractor assembly,4. Insert the shaft of the Cloward set into the ⅜″ variable speed drilland tighten the chuck with the key. The set-point is then placed andlocked at the forward aspect of the cutter.

[0122] The apparatus is then placed on the tissue to be fashioned.Drilling is commenced at a moderate speed. After the set-point has madea deep cut in the tissue, and the teeth have begun to cut into thetissue, drilling is stopped, and the set-point apparatus is unlocked.Drilling is continued using the marks created as a guide.

[0123] The Cloward(s) are then removed from the tissue block. A Stryker®saw or band saw is then used to remove the cut grafts after all havebeen cut. Any cartilage is then trimmed from the cortical face of theCloward(s) using a scalpel and a # 10 blade. The distal end of the graftis then trimmed perpendicular to the body of the graft with a band sawmaking sure the fashioned graft is at least 15 mm long. The Cloward(s)are cleansed using pulsatile water apparatus. If the surface marrow isnot easily removed, dry spin the graft(s) at 2600 rpm for 3 minutes.

[0124] The Cloward(s) are then placed in a sterile container withhydrogen peroxide (3%) at 37 to 44° C. The container is sealed and thecontainer is placed into the centrifuge. The centrifuge is thenbalanced. The grafts are then centrifuged at 2600 rpm for 15 minutes.The tissue is removed from the centrifuge and the grafts are placed intoan ultrasonic cleaner. Equal volumes of Allowash™ Solution, hydrogenperoxide (3%), and antibiotics are added to the ultrasonic cleaner andsonicate the tissue at 37-44° C. for a minimum of 1 hour. Thereafter,the tissue is removed from the ultrasonic cleaner.

[0125] The mixture is decanted and a sterile glass container is filledwith fresh 3% hydrogen peroxide. The grafts are then placed in thecontainer, the top is sealed and the container is taken to the largeultrasonic cleaner. The grafts are then sonicated for 90 minutes.Thereafter, the grafts are incubated overnight at 37-44° C. (minimum of6 hours, preferably 12 to 18 hours).

[0126] After incubation, the hydrogen peroxide is decanted and the basinis filled with 70% isopropyl alcohol and the grafts are incubated atroom temperature for a minimum of 30 minutes. Thereafter, the isopropylalcohol is decanted and the container is filled with warm sterile water.The grafts are incubated for a minimum of 30 minutes. Methods ofincubation include for example: soaking and mild agitation.

[0127] The wash solution is then decanted and the Cloward dowels areremoved from the container. The Cloward dowels are then placed into asterile container. The container is sealed and placed into thecentrifuge. The centrifuge is balanced. The grafts are then centrifugedfor 3-5 minutes to dry and the remaining solution is removed.

[0128] The width and length of the Cloward(s) are measured, graftidentification numbers are assigned, and the information is recorded onthe “Tissue Processing Log Worksheet”. One graft is then placed into aglass, 120 cc bottle and the printed label is affixed with the uniquenumeric designator. This step is repeated until all deposits arebottled. The bottled grafts are either frozen and packaged, or frozenand freeze-dried and packaged.

[0129] B. Plasticization:

[0130] Viscous glycerol is then added to each bottle sufficient to coverthe graft and vacuum (10 to 500 mTorr) is applied to each bottle untilthe air ceases to exit the grafts (usually 5-20 minutes depending ongraft type). The grafts are then removed from the bottles and placedinto a centrifuge container. The grafts are centrifuged for 15-30minutes or until glycerol ceases to exit the grafts and accumulate inthe space below the grafts. The bottled grafts are either packaged orplaced under vacuum and packaged.

EXAMPLE 7 Processing Cloward Dowels

[0131] A. Cleaning and Processing:

[0132] Graft material is selected and all of the soft tissue andperiosteum is removed from the distal femur, proximal and distal tibia,and cartilage is removed from the site. The femur is transected 10-15 cmabove the femoral condyles and the distal femoral condyles are bisected.Transect the proximal tibia 10-15 cm below the tibial plateau. Thedistal femur or proximal tibia is placed in a Pan-A-Vise™. This isaccomplished by removing a section of the diaphysis, allowing the visejaws to grip the tissue securely. The Cloward set (12, 14, 16, 18, or 20mm) is then assembled: 1. Place the extractor assembly within the cuttershaft, 2. Screw the cutter assembly onto the shaft with the aid of theCloward set wrench, 3. Screw the set-point onto the extractor assembly,4. Insert the shaft of the Cloward set into the ⅜″ variable speed drilland tighten the chuck with the key. The set-point is then placed andlocked at the forward aspect of the cutter.

[0133] The apparatus is then placed on the tissue to be fashioned.Drilling is commenced at a moderate speed. After the set-point has madea deep cut in the tissue, and the teeth have begun to cut into thetissue, drilling is stopped, and the set-point apparatus is unlocked.Drilling is continued using the marks created as a guide.

[0134] The Cloward(s) are then removed from the tissue block. A Stryker®saw or band saw is then used to remove the cut grafts after all havebeen cut. Any cartilage is then trimmed from the cortical face of theCloward(s) using a scalpel and a # 10 blade. The distal end of the graftis then trimmed perpendicular to the body of the graft with a band sawmaking sure the fashioned graft is at least 15 mm long. The Cloward(s)are cleansed using pulsatile water apparatus. If the surface marrow isnot easily removed, dry spin the graft(s) at 2600 rpm for 3 minutes.

[0135] B. Plasticization:

[0136] The Cloward(s) are then placed in a sterile container withhydrogen peroxide (3%)/glycerin USP 30% at 37 to 44° C. The container issealed and the container is placed into the centrifuge. The centrifugeis then balanced. The grafts are then centrifuged at 2600 rpm for 15minutes. The tissue is removed from the centrifuge and the grafts areplaced into an ultrasonic cleaner. Equal volumes of Allowash™ Solution,hydrogen peroxide (3%), and antibiotics are added to the ultrasoniccleaner and sonicate the tissue at 37-44° C. for a minimum of 1 hour.Thereafter, the tissue is removed from the ultrasonic cleaner.

[0137] The mixture is decanted and a sterile glass container is filledwith fresh 3% hydrogen peroxide. The grafts are then placed in thecontainer, the top is sealed and the container is taken to the largeultrasonic cleaner. The grafts are then sonicated for 90 minutes.Thereafter, the grafts are incubated overnight at 37-44° C. (minimum of6 hours, preferably 12 to 18 hours).

[0138] After incubation, the hydrogen peroxide is decanted and the basinis filled with 70% isopropyl alcohol/30% glycerin USP and the grafts areincubated at room temperature for a minimum of 30 minutes. Thereafter,the isopropyl alcohol/glycerin USP solution is decanted and thecontainer is filled with warm 30% glycerin USP in sterile water. Thegrafts are incubated for a minimum of 30 minutes. Methods of incubationinclude for example: soaking and mild agitation.

[0139] The solution is then decanted and the Cloward dowels are removedfrom the container. The Cloward dowels are then placed into a sterilecontainer. The container is sealed and placed into the centrifuge. Thecentrifuge is balanced. The grafts are then centrifuged for 3-5 minutesto dry and the remaining solution is removed.

[0140] The width and length of the Cloward(s) are measured, graftidentification numbers are assigned, and the information is recorded onthe “Tissue Processing Log Worksheet”. One graft is then placed into aglass, 120 cc bottle and the printed label is affixed with the uniquenumeric designator. This step is repeated until all deposits arebottled. The bottled grafts are either frozen and packaged, or frozenand freeze-dried and packaged.

EXAMPLE 8 Processing of an Iliac Crest Wedge

[0141] A. Cleaning and Processing:

[0142] The soft tissue, periosteum, and cartilage is removed from anilium. The ilium is placed in a Pan-A-Vise™ by removing a section of theilium, allowing the vise jaws to grip the tissue securely. A Stryker sawis assembled with parallel cutting blades (12, 14, 16, 18, or 20 mm).The set-point at the forward aspect of the cutter is placed and locked.The apparatus is placed on the tissue to be fashioned and cutting isbegun at a moderate speed.

[0143] After the set-point has made a deep cut in the tissue, and theteeth have begun to cut into the tissue, cutting is stopped, and theset-point apparatus is checked. Cutting is continued using the markscreated as a guide. A Stryker® saw or band saw is then used to removethe cut grafts after all have been cut.

[0144] Any cartilage is trimmed from the cortical face of the grafts(s)using a scalpel and a # 10 blade. The distal end of the graftperpendicular to the body of the graft is trimmed with a band saw makingsure the fashioned graft is at least 15 mm long. The Grafts is thencleansed using a pulsatile water apparatus. If the surface marrow is noteasily removed, the graft(s) is dry spun at 2600 rpm for 3 minutes.

[0145] B. Plasticization:

[0146] The Iliac Crest Wedge(s) are then placed in a sterile containerwith hydrogen peroxide (3%) and glycerin USP (30%) at 37 to 44° C. Thecontainer is sealed and placed into the centrifuge. The centrifuge isbalanced. The grafts are then centrifuged at 2600 rpm for 15 minutes.The tissue is removed from the centrifuge and the grafts are placed intothe ultrasonic cleaner. Equal volumes of Allowash™ Solution, hydrogenperoxide (3%), glycerin USP (30%), and antibiotics are added to theultrasonic cleaner, and the grafts are sonicated at 37-44° C. for aminimum of 1 hour.

[0147] The tissue is then removed from the ultrasonic cleaner. Themixture is decanted and a sterile glass container is filled with fresh3% hydrogen peroxide/30% glycerin USP. The grafts are placed in thecontainer, the top is sealed and the container is taken to a largeultrasonic cleaner. The grafts are sonicated for 90 minutes. Thereafter,the grafts are incubated overnight at 37-44° C. (minimum of 6 hours,preferably for 12 to 18 hours). Methods of incubation include forexample: soaking and mild agitation.

[0148] The hydrogen peroxide/glycerin USP is then decanted and the basinis filled with 70% isopropyl alcohol/30% glycerin USP. The grafts arethen incubated at room temperature for a minimum of 30 minutes. Theisopropyl alcohol/glycerin USP solution is then decanted and thecontainer is filled with warm 30% glycerin USP in water. The grafts areincubated for a minimum of 30 minutes.

[0149] The glycerin USP solution is then decanted and the Iliac CrestWedges are removed from the container. The Iliac Crest Wedges are thenplaced into a sterile container. The container is sealed and placed intothe centrifuge. The centrifuge is balanced and the grafts arecentrifuged for 3-5 minutes to dry and the remaining solution isremoved.

[0150] The width and length of the Wedges are measured, graftidentification numbers are assigned, and the information is recorded onthe “Tissue Processing Log Worksheet”. One graft is then placed into aglass, 120 cc bottle and the printed label is affixed with the uniquenumeric designator. This step is repeated until all deposits arebottled. The bottled grafts are either frozen and packaged, or frozenand freeze-dried and packaged.

EXAMPLE 9 Processing of Fascia Lata

[0151] A. Cleaning and Processing:

[0152] Any remaining muscle tissue is removed from the fascia lata. Thefascia is placed with the subcutaneous layer uppermost, on a clean,drape towel. Using blunt dissection techniques, all of the fat andextraneous soft tissue is removed from the graft material. The graft iskept moist with sterile water to prevent desiccation during processing.

[0153] Any torn fibers are removed from the edges of the graft material,and a graft rectangular in shape is created. The graft(s) are thenplaced in a basin containing a 1:100 dilution of Allowash™ Solution orother surfactant(s) for at least 15 minutes. The basin is labeled asAllowash™ Solution. The time of exposure is recorded on the TissueProcessing Log Worksheet.

[0154] B. Plasticization:

[0155] The graft(s)are placed into an empty basin labeled “Rinse”. Thegraft(s) are rinsed three time with copious amounts of sterile water toremove any residual detergents. Any sterile water which accumulates inthe Rinse basin is discarded. The number of rinses is recorded on theTissue Processing Log Worksheet. The fashioned graft(s) are then placedin the basin containing U.S.P. grade 70% isopropyl alcohol containing30% glycerin USP for 2-5 minutes. The basin is labeled IPA/Glycerin. Thetime of exposure to the alcohol/glycerin USP solution is recorded in theTissue Processing Log Worksheet.

[0156] The graft(s) are then placed into the basin containing theantibiotic solution in 30% glycerin USP for at least 15 minutes. Thebasin is labeled as Antibiotics/Glycerin USP. The exposure time to theantibiotics/glycerin USP is recorded on the Tissue Processing LogWorksheet. The graft(s) are then thoroughly soaked by immersing eachdeposit into sterile 30% glycerin USP in deionized/distilled water for aminimum of 5 minutes to remove excess antibiotics. Enough sterileglycerin USP solution is needed to cover the graft(s). The basin islabeled as Rinse. The time of exposure to the glycerin USP rinsesolution is recorded on the Tissue Processing Log Worksheet.

[0157] The fashioned graft(s) are then placed on sterile fine meshgauze, and the gauze is trimmed to just beyond the edges of the graft.The width and length of the graft(s) is measured to the nearest tenth ofa centimeter. The graft(s) are assigned identification numbers and thisinformation is recorded on the Tissue Processing Log Worksheet. Thegraft and gauze is then rolled into a tube and graft material is thenplaced into glass, 120 ml bottles, and the printed label is affixed withthe unique numeric designator. This step is repeated until all depositsare bottled. The graft material is now ready for wrapping andfreeze-drying or dehydrating.

EXAMPLE 10 Processing Pericardium

[0158] A. Cleaning and Processing:

[0159] The pericardial tissue is rinsed of any blood or pericardialfluid in sterile water in the basin labeled Rinse. The pericardium isthen placed on a clean drape towel. Using blunt dissection techniques,all of the fat and extraneous soft tissue is removed from the graftmaterial. The graft is kept moist with sterile water to preventdesiccation during processing. Any torn fibers are removed from theedges of the graft material, and a graft rectangular in shape iscreated. The graft(s) are then placed in a basin containing a 1:100dilution of Allowash Solution or other surfactant(s) for at least 15minutes. The basin is labeled as Allowash™ Solution. The time ofexposure is recorded on the Tissue Processing Log Worksheet.

[0160] The graft(s)are placed into an empty basin labeled “Rinse”. Thegraft(s) are rinsed three time with copious amounts of sterile water toremove any residual detergents. Any sterile water which accumulates inthe Rinse basin is discarded. The number of rinses is recorded on theTissue Processing Log Worksheet. The fashioned graft(s) are then placedin the basin containing U.S.P. grade 70% isopropyl alcohol containing30% glycerin USP for 2-5 minutes. The basin is labeled IPA/Glycerin. Thetime of exposure to the alcohol/glycerin USP solution is recorded in theTissue Processing Log Worksheet.

[0161] The graft(s) are then placed into the basin containing theantibiotic solution in 30% glycerin USP for at least 15 minutes. Thebasin is labeled as Antibiotics/Glycerin USP. The exposure time to theantibiotics/glycerin USP is recorded on the Tissue Processing LogWorksheet. The graft(s) are then thoroughly soaked by immersing eachdeposit into sterile 30% glycerin USP in deionized/distilled water for aminimum of 5 minutes, preferably from 10 to 15 minutes, to remove excessantibiotics. Enough sterile glycerin USP solution is needed to cover thegraft(s). The basin is labeled as Rinse. The time of exposure to theglycerin USP rinse solution is recorded on the Tissue Processing LogWorksheet.

[0162] The fashioned graft(s) are then placed on sterile fine meshgauze, and the gauze is trimmed to just beyond the edges of the graft.The width and length of the graft(s) is measured to the nearest tenth ofa centimeter. The graft(s) are assigned identification numbers and thisinformation is recorded on the Tissue Processing Log Worksheet. Thegraft and gauze is then rolled into a tube and graft material is thenplaced into glass, 120 ml bottles, and the printed label is affixed withthe unique numeric designator. This step is repeated until all depositsare bottled. The graft material is now wrapped and placed in a freezedryer or dehydrated.

[0163] All of the publications and patent applications cited herein arehereby incorporated by reference into the present disclosure. It will beappreciated by those skilled in the art that various modifications canbe made without departing from the essential nature thereof It isintended to encompass all such modifications within the scope of theappended claims.

What is claimed:
 1. A plasticized bone graft, comprising: a cleaned bone graft comprising one or more plasticizers.
 2. A plasticized bone graft suitable for transplantation into a human, comprising: a bone graft having an internal matrix essentially free from bone marrow elements; and one or more plasticizers contained in said internal matrix.
 3. A plasticized bone graft, comprising: a cleaned bone graft; and one or more plasticizers, wherein said cleaned bone graft is impregnated with said one or more plasticizers.
 4. A method for producing a plasticized bone graft suitable for transplantation into a human, comprising: impregnating a cleaned bone graft with one or more plasticizers to produce a plasticized bone graft.
 5. The method of claim 4 , said step of impregnating, comprising: incubating said cleaned bone graft with a plasticizer composition comprising one or more plasticizers and one or more biocompatible solvents.
 6. The method of claim 5 , wherein said one or more biocompatible solvents comprise one or more alcohols.
 7. The method of claim 5 , wherein incubating comprises soaking said cleaned bone graft in said plasticizer composition.
 8. The method of claim 7 , wherein said soaking is carried out under negative pressure.
 9. The bone graft of any one of claims 1, 2, or 3, wherein said bone graft is suitable for direct transplant into a human without rehydration.
 10. The method of claim 4 , wherein said cleaned bone graft is essentially free from bone marrow elements.
 11. The method of claim 6 , wherein said one or more plasticizers are present in said plasticizer composition in a weight ratio of from 30 to 90 wt %, and said one or more alcohols are present in said plasticizer composition in a weight ratio of from 10 to 70 wt %.
 12. The method of claim 11 , wherein said plasticizer is glycerol and said alcohol is isopropyl alcohol.
 13. The method of claim 12 , wherein said glycerol is present at 30 wt % and said isopropyl alcohol is present at 70 wt %.
 14. The bone graft of any one of claims 1, 2, or 3, wherein said bone graft is a non-demineralized bone graft.
 15. The bone graft of any one of claims 1, 2, or 3, wherein said bone graft is a load-bearing bone graft.
 16. A plasticized soft tissue graft suitable for transplantation into a human, comprising: a cleaned soft tissue graft having an internal matrix; and one or more plasticizers contained in said internal matrix.
 17. A plasticized soft tissue graft, comprising: a cleaned, soft tissue graft; and one or more plasticizers, wherein said cleaned soft tissue graft is impregnated with said one or more plasticizers.
 18. A plasticized soft tissue graft, comprising: a cleaned, soft tissue graft comprising one or more plasticizers.
 19. A method for producing a plasticized soft tissue graft suitable for transplantation into a human, comprising: impregnating a cleaned, soft tissue graft with one or more plasticizers to produce a plasticized soft tissue graft.
 20. The method of claim 19 , said step of impregnating, comprising: incubating said cleaned, soft tissue graft with a plasticizer composition comprising one or more plasticizers and one or more biocompatible solvents.
 21. The method of claim 20 , wherein said one or more biocompatible solvents comprise one or more alcohols.
 22. The method of claim 20 , wherein incubating comprises soaking said cleaned, soft tissue graft in said plasticizer composition.
 23. The method of claim 22 , wherein said soaking is carried out under negative pressure.
 24. The soft tissue graft of any one of claims 16, 17, or 18, wherein said soft tissue graft is suitable for direct transplant into a human without rehydration.
 25. The method of claim 21 , wherein said one or more plasticizers are present in said plasticizer composition in a weight ratio of from 30 to 90 wt %, and said one or more alcohols are present in said plasticizer composition in a weight ratio of from 10 to 70 wt %.
 26. The method of claim 25 , wherein said plasticizer is glycerol and said alcohol is isopropyl alcohol.
 27. The method of claim 26 , wherein said glycerol is present at 30 wt % and said isopropyl alcohol is present at 70 wt %.
 28. The soft tissue graft of any one of claims 16, 17, or 18, wherein said soft tissue graft is a load-bearing soft tissue graft.
 29. The soft tissue graft of any one of claims 16, 17, or 18, wherein said soft tissue graft is selected from the group consisting of: dura, pericardium, fascia lata, tendons and ligaments
 30. A plasticized bone graft, comprising: a cleaned, non-demineralized bone graft comprising one or more plasticizers.
 31. A plasticized load-bearing bone graft, comprising: a cleaned, load-bearing bone graft comprising one or more plasticizers.
 32. A plasticized load-bearing soft tissue graft, comprising: a cleaned, load-bearing soft tissue graft comprising one or more plasticizers. 